BIOCHEMICAL AND CYTOGENEITC MARKERS IN RETINOBLASTOMA

这项建议的长期目标是了解 视网膜母细胞瘤(Rb)基因，即Rb功能丧失 人类肿瘤的发展及其作为预后指标的潜力 肿瘤患者临床转归的指标 已发生RB功能丢失。初步研究将集中在 如肺癌、乳腺癌、膀胱癌、骨癌、卵巢癌和 肾脏和急性髓系白血病。只要有可能就正常 来自同一部位的组织将被检查，转移性病变也将被检查 如果有的话，取自同一个病人。 主要重点将放在检查人类肿瘤的RB状态 冰冻切片和石蜡切片中的单细胞水平以及 直接免疫印迹，尽管许多肿瘤将是 此外，还在RNA和DNA水平上进行了检查。后者将会完成 在我们试图找出发生在 Rb基因引起的功能丧失与确认和补体 我们的蛋白质研究。石蜡切片的使用将在任何时候 有可能，因为我们最近成功地获得了优秀的 石蜡切片的免疫组织化学Rb染色 多克隆Rb抗体，据我们所知，这是不可能的 任何商业上可获得的Rb抗体，无论是多克隆还是 单克隆性。 这项提案的目标的实现将提供更多的 更好地理解Rb基因在一些最 人类常见的恶性肿瘤，同样或更重要的是 确定是否监测特定疾病患者的RB状态 恶性肿瘤将提供与其相关的重要信息 临床结果或疾病的侵袭性。

The long term objective of this proposal is to understand the role of the retinoblastoma (RB) gene, namely loss of RB function, in the development of human tumors as well as its potential to be a prognostic indicator of the clinical outcome in patients with tumors in which the loss of RB function has occurred. Initial studies will concentrate on such tumors as cancers of the lung, breast, bladder, bone, ovary and kidney as well as acute myeloblastic leukemia. Whenever possible normal tissue from the same site will be examined as will metastatic lesions from the same patient when available. Major emphasis will be placed on examining the RB status of human tumors at the single cell level in frozen and paraffin sections as well as direct Western immunoblotting, although many of the tumors will be examined in addition at the RNA and DNA levels. The latter will be done in our attempt to find the structural changes that have occurred within the RB gene to cause the loss of function and to confirm and complement our protein studies. The use of paraffin sections will be done whenever possible since we have recently been successful in obtaining excellent immunohistochemical RB staining of paraffin sections using our polyclonal RB antibody, which, to our knowledge, is not possible with any commercially available RB antibody, whether polyclonal or monoclonal. The fulfillment of the goals of this proposal should provide a much better understanding of the role of the RB gene in some of the most common human malignancies and equally or perhaps even more importantly determine whether monitoring the RB status in patients with specific malignancies will provide significant information related to their clinical outcome or the aggressiveness of their disease.